Hiemer B, Genz B, Jonitz-Heincke A, Pasold J, Wree A, Dommerich S, Bader R
Rostock University Medical Center, Department of Orthopaedics, Biomechanics and Implant Technology Research Laboratory, Doberaner Strasse 142, 18057 Rostock, Germany.
Rostock University Medical Center, Department of Otorhinolaryngology, Doberaner Strasse 137-139, 18057 Rostock, Germany.
Sci Rep. 2016 Sep 27;6:33747. doi: 10.1038/srep33747.
The regeneration of cartilage lesions still represents a major challenge. Cartilage has a tissue-specific architecture, complicating recreation by synthetic biomaterials. A novel approach for reconstruction is the use of devitalised cartilage. Treatment with high hydrostatic pressure (HHP) achieves devitalisation while biomechanical properties are remained. Therefore, in the present study, cartilage was devitalised using HHP treatment and the potential for revitalisation with chondrocytes and mesenchymal stem cells (MSCs) was investigated. The devitalisation of cartilage was performed by application of 480 MPa over 10 minutes. Effective cellular inactivation was demonstrated by the trypan blue exclusion test and DNA quantification. Histology and electron microscopy examinations showed undamaged cartilage structure after HHP treatment. For revitalisation chondrocytes and MSCs were cultured on devitalised cartilage without supplementation of chondrogenic growth factors. Both chondrocytes and MSCs significantly increased expression of cartilage-specific genes. ECM stainings showed neocartilage-like structure with positive AZAN staining as well as collagen type II and aggrecan deposition after three weeks of cultivation. Our results showed that HHP treatment caused devitalisation of cartilage tissue. ECM proteins were not influenced, thus, providing a scaffold for chondrogenic differentiation of MSCs and chondrocytes. Therefore, using HHP-treated tissue might be a promising approach for cartilage repair.
软骨损伤的修复仍然是一项重大挑战。软骨具有特定的组织结构,这使得用合成生物材料进行再造变得复杂。一种新的重建方法是使用失活软骨。高静水压(HHP)处理可实现软骨失活,同时保留其生物力学性能。因此,在本研究中,使用HHP处理使软骨失活,并研究了软骨细胞和间充质干细胞(MSCs)使其恢复活力的潜力。通过在10分钟内施加480MPa的压力来实现软骨的失活。通过台盼蓝排斥试验和DNA定量证明了有效的细胞失活。组织学和电子显微镜检查显示HHP处理后软骨结构未受损。为了实现恢复活力,将软骨细胞和MSCs培养在失活的软骨上,不添加软骨生成生长因子。软骨细胞和MSCs均显著增加了软骨特异性基因的表达。细胞外基质(ECM)染色显示,培养三周后出现了类似新软骨的结构,阿赞染色呈阳性,同时有II型胶原蛋白和聚集蛋白聚糖沉积。我们的结果表明,HHP处理导致软骨组织失活。ECM蛋白未受影响,因此为MSCs和软骨细胞的软骨分化提供了支架。因此,使用HHP处理的组织可能是一种有前景的软骨修复方法。